Antioksidan Etkili Rutin İçeren Mikroemülsiyon ve Mikroemülsiyon Jel Formülasyonlarının Geliştirilmesi, Karakterizasyonu ve Etkinliğinin İncelenmesi

Öz

Amaç: Çalışmada antioksidan etkili doğal bileşik olan rutinin, mikroemülsiyon (ME) ve mikroemülsiyon jel (MEj) formülasyonlarının geliştirilmesi ve uzatılmış antioksidan etkinliğinin değerlendirilmesi amaçlanmıştır. Gereç ve Yöntemler: Farklı oranlarda yağ/su/yüzey etkin madde:yardımcı yüzey etkin madde kullanılarak hazırlanan ME formülasyonlarının damlacık büyüklüklükleri, zeta potansiyel değerleri, viskoziteleri ve bekletme, santrifüj ve dondurup çözme işlemleri sonrasındaki stabiliteleri tayin edilmiştir. Altı farklı ME formülasyonu arasından belirlenen stabil formülasyonlar ile MEj formülasyonları, %1’lik (a/h) Karbopol 938 kullanılarak hazırlanmıştır. Karakterizasyon çalışmaları yanı sıra L929 fare fibroblast hücre hattı kullanılarak hücre kültüründe toksisite testi, in vitro salım ve hücre geçiş çalışmaları yapılmıştır. Veriler ışığında en iyi belirlenen formülasyonun (F2 ME) antioksidan aktivitesi, 1,1-difenil-2-pikrilhidrazil (DPPH●) ve (2,2-Azino-bis3-etilbenzotiyazolin-6-sülfonik asit) (ABTS●+) radikal süpürücü etki ile belirlenmiştir. Bulgular: Altı farklı mikroemülsiyon formülasyonu geliştirilmiş ve karakterize edilmiştir. Toksisite çalışmasına göre, rutin çözeltisi ve rutin ile geliştirilen formülasyonlar toksik bulunmamıştır. Karakterizasyon, in vitro salım ve hücre geçiş çalışması verileri ile en iyi formülasyon olarak belirlenen F2 kodlu ME formülasyonundan salınan rutin miktarının antioksidan etkisi incelenmiş ve DPPH● ve ABTS+● radikallerini süpürücü etkisi ispatlanmıştır. Sonuç: Sonuç olarak, bu çalışma ile toksik etkisi bulunmayan, yeni, rutin yüklü ME ve MEj formülasyonları başarıyla geliştirilmiş, uzatılmış antioksidan etkinlik kanıtlanmıştır. Bu çalışmada geliştirilen rutin yüklü ME formülasyonunun topikal uygulama için potansiyel oluşturduğu düşünülmektedir.

Anahtar Kelimeler

• rutin
• mikroemülsiyon
• mikroemülsiyon jel
• antioksidanlar

Development, Characterization and Evaluation of Effectiveness of Microemulsion and Microemulsion Gel Formulations Containing Antioxidant Effective Rutin

Öz

im: In this study, it was aimed to develop microemulsion and microemulsion gel formulations of a natural product (rutin) that possesses antioxidant effect and to evaluate its prolonged antioxidant activity. Material and Methods: The droplet size, zeta potential, viscosity values and stability profiles following the holding, centrifugation and freeze-thawing processes applied to the ME formulations, prepared by using different ratios of oil/water/surfactant and co-surfactant, were determined. The most stable formulations out of the six different ME formulations were selected and then, the ME gel formulations were prepared using 1% (w/v) Carbopol 938. Besides characterization studies, toxicity test on cell culture with using L929 mouse fibroblast cell line, in vitro release and cell permeation studies were performed. Antioxidant activity of optimum formulation (F2 coded ME) were measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH) ve 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS●+) radical scavenging effect. Results: Six different ME formulations were developed and characterized. According to the toxicity study, rutin solution and rutin loaded formulations were not toxic. The antioxidant effect of the rutin released from the F2 coded ME formulation which was determined to be optimum formulation by characterization, in vitro release and cell permeation studies data was investigated and the scavenging effects of DPPH and ABTS + radicals were proven. Conclusion: In conclusion, new, nontoxic, rutin loaded ME and ME gel formulations were successfully developed and prolonged antioxidant activity was proved. It is assumed that the rutin loaded ME formulation developed in this study has a great potential for topical application.

Anahtar Kelimeler

• Rutin
• microemulsion
• microemulsion gel
• antioxidants

Kaynakça

1. Talegaonkar S, Azeem A, Ahmad F, Khar R, Pathan S, Khan Z. Microemulsions: a novel approach to enhanced drug delivery. Recent Patents on Drug Delivery & Formulation. 2008; 2(3): 238-57.
2. Katiyar BS, Katiyar SS, Mishra PS, Sailaja DL. Microemulsions: a novel drug carrier system. International Journal of Pharmaceutical Sciences Review and Research. 2013; 20(2): 138-48.
3. Ezrahi S, Aserin A, Garti N. Basic principles of drug delivery systems - the case of paclitaxel. Adv Colloid Interface Sci. 2019; 263: 95-130.
4. Schulman JH, Stoeckenius W, Prince LM. Mechanism of formation and structure of micro emulsions by electron microscopy. The Journal of Physical Chemistry. 1959; 63(10): 1677-80.
5. Lopes LB. Overcoming the cutaneous barrier with microemulsions. Pharmaceutics. 2014; 6(1): 52-77.
6. Hajjar B, Zier K-I, Khalid N, Azarmi S, Löbenberg R. Evaluation of a microemulsion-based gel formulation for topical drug delivery of diclofenac sodium. Journal of Pharmaceutical Investigation. 2017; 48(3): 351-62.
7. Morishita T, Hajika M, Sakai S, Tetsuka T. Development of simple spectrophotometric assay for the rutin-degrading enzyme in buckwheat. Current Advances in Buckwheat Research. 1995; 6: 833-7.
8. Saklani R, Gupta SK, Mohanty IR, Kumar B, Srivastava S, Mathur R. Cardioprotective effects of rutin via alteration in TNF-alpha, CRP, and BNP levels coupled with antioxidant effect in STZ-induced diabetic rats. Molecular and Cellular Biochemistry. 2016; 420(1-2): 65-72.

9. Chen H, Chang X, Du D, Li J, Xu H, Yang X. Microemulsion-based hydrogel formulation of ibuprofen for topical delivery. International Journal of Pharmaceutics. 2006; 315(1-2): 52-8.
10. Mali KK, Dhawale SC, Dias RJ. Microemulsion based bioadhesive gel of itraconazole using tamarind gum: in-vitro and ex-vivo evaluation. Marmara Pharmaceutical Journal. 2017; 21(3): 688-700.
11. Sumedha P, Narayana CR, Sandeep D, Ravi G, Anoop NV. Microemulsion based gel of sulconazole nitrate for topical application. Turkish Journal of Pharmaceutical Sciences. 2020; 17(3): 259-64.
12. Khalil Y, Khasraghi A, Mohammed E. Preparation and evaluation of physical and, rheological properties of clotrimazole emulgel. Iraqi Journal of Pharmaceutical Sciences. 2011; 20(2): 19-27.
13. Yücel Ç, Şeker Karatoprak G, Değim IT. Anti-aging formulation of rosmarinic acid-loaded ethosomes and liposomes. Journal of Microencapsulation. 2019; 36(2): 180-91.
14. Gyamfi M, Yonamine M, Aniya Y. Free-radical scavenging action of medicinal herbs from Ghana: thonningia sanguinea on experimentally induced liver injuries. General Pharmacology. 1999; 32(6): 661-7.
15. Re R, Pellegrini NP, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology & Medicine. 1999; 26(9-10): 1231-7.
16. Abualhasan MN, Mansour J, Jaradat N, Zaid AN, Khadra I. Formulation and development of a validated UV-spectrophotometric analytical method of rutin tablet. Int Sch Res Notices. 2017; 2017: 1-7. doi: 10.1155/2017/2624947
17. Ahmad M, Sahabjada -, Akhtar J, Hussain A, Badaruddeen -, Arshad M, et. al. Development of a new rutin nanoemulsion and its application on prostate carcinoma PC3 cell line. EXCLI Journal. 2017; 16: 810-23.
18. Savale SK. Formulation and evaluation of intranasal microemulsion containing rutin. Asian Journal of Biomaterial Research. 2017; 3(6): 25-7.
19. Pate K, Safier P. Chemical metrology methods for CMP quality. In: Babu S, editor. Advances in chemical mechanical planarization (CMP). Newyork: Woodhead Publishing; 2016. p. 299-325.
20. Songkro S, Lo NL, Tanmanee N, Maneenuan D, Boonme P. In vitro release, skin permeation and retention of benzophenone-3 from microemulsions (o/w and w/o). Journal of Drug Delivery Science and Technology. 2014; 24(6): 703-11.
21. Kajbafvala A, Salabat A, Salimi A. Formulation, characterization, and in vitro/ex vivo evaluation of quercetin-loaded microemulsion for topical application. Pharm Dev Technol. 2018; 23(8): 741-50.
22. Ingels F, Deferme S, Destexhe E, Oth M, Van den Mooter G, Augustijns P. Simulated intestinal fluid as transport medium in the Caco-2 cell culture model. International Journal of Pharmaceutics. 2002; 232(1-2): 183-92.
23. Cui Y. Investigation of microemulsion system for transdermal delivery of ligustrazine phosphate. African Journal of Pharmacy and Pharmacology. 2011; 5(14): 1674-81.